Translational RelevanceThere is an urgent need for progress in the management of breast cancer leptomeningeal metastasis (BCLM). Current diagnostics are hampered by impaired sensitivity, delaying diagnosis and treatment initiation. Further, during BCLM therapy there are no quantitative response markers to guide clinical decision making. This proof-of-concept study explored the use of ulpWGS, a methodology requiring no upfront knowledge of tumor mutations, to detect ctDNA in cerebrospinal fluid (CSF) in BCLM. ctDNA was detectable in all BCLM+ patients by ulpWGS, despite frequent negative CSF cytology. Importantly, CSF ctDNA was not detected in BCLM-free patients, supporting the potential for ulpWGS as a tool to refute diagnosis. In addition, CSF ctDNA reduction, measured by ulpWGS, was associated with improved survival on intrathecal BCLM treatment. This study highlights the potential of ulpWGSassessed CSF ctDNA fraction to improve BCLM care through timely diagnosis and adaptation of therapy, and warrants larger prospective studies to test clinical validity.